title: A novel and simple method for generation of human dendritic cells from unfractionated peripheral blood mononuclear cells within 2 days: its application for induction of HIV-1-reactive CD4(+) T cells in the hu-PBL SCID mice.
abstract: Because dendritic cells (DCs) play a critical role in the regulation of adaptive immune responses, they have been ideal candidates for cell-based immunotherapy of cancers and infections in humans. Generally, monocyte-derived DCs (MDDCs) were generated from purified monocytes by multiple steps of time-consuming physical manipulations for an extended period cultivation. In this study, we developed a novel, simple and rapid method for the generation of type-1 helper T cell (Th1)-stimulating human DCs directly from bulk peripheral blood mononuclear cells (PBMCs). PBMCs were cultivated in the presence of 20 ng/ml of granulocyte-macrophage colony-stimulating factor, 20 ng/ml of interleukin-4 (IL-4) and 1,000 U/ml of interferon-β for 24 h followed by 24 h maturation with a cytokine cocktail containing 10 ng/ml of tumor necrosis factor-α (TNF-α), 10 ng/ml of IL-1β and 1 μg/ml of prostaglandin E2. The phenotype and biological activity of these new DCs for induction of allogeneic T cell proliferation and cytokine production were comparable to those of the MDDCs. Importantly, these new DCs pulsed with inactivated HIV-1 could generated HIV-1-reactive CD4(+) T cell responses in humanized mice reconstituted with autologous PBMCs from HIV-1-negative donors. This simple and quick method for generation of functional DCs will be useful for future studies on DC-mediated immunotherapies.
authors.affiliation: Department of Immunology, Graduate School of Medicine, University of the Ryukyus Okinawa, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Kodama
authors.forename: Reiko
authors.initials: R
authors.lastname: Tanaka
authors.forename: Mineki
authors.initials: M
authors.lastname: Saito
authors.forename: Aftab A
authors.initials: AA
authors.lastname: Ansari
authors.forename: Yuetsu
authors.initials: Y
authors.lastname: Tanaka
keywords: Th1-inducing DCs; anti-HIV-1 T cell response; dendritic cell; hu-PBL-SCID; short-term culture